Restriction fragment length polymorphism (RFLP) analysis was performed by digesting the phage DNA with the enzymes HhaI and MspI (NEB) according to the manufacturer's instructions. Electron Microscopy. Kanamycin stock solution is sterilized by filtration using 0.22 . Phage DNA was extracted using a phage DNA isolation kit (Norgen Biotek, Ontario, Canada). For next-generation sequencing of the phage PLG-II genome, a 150-bp paired-end DNA fragment library was prepared using a Nextera XT DNA Library Prep Kit (Illumina, Inc., USA) according to the manufacturer's protocol. M13 Phage DNA Rapid Extraction Kit-new revised Author: Fausto Created Date: 9/29/2010 3:16:02 PM Keywords () . The quantity and quality of DNA for sequencing were assessed both on BioDrop (biochrom Ltd., Cambridge, UK) and Qubit3.0 fluorometer . Effective Host Genomic DNA Removal without Reducing Phage DNA Yield. Phage dilution buffer and media must be autoclaved prior to use and stored at 25C. US EN. Bacteriophage DNA was extracted using the Phage DNA isolation kit (Norgen Biotek Corporation), and was sequenced, assembled and annotated as previously described (Esplin et al., 2017; Sharma et al., 2018). Norgen's Phage DNA Isolation Kit is a quick method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. For genome sequencing, phage nucleic acid was extracted using a Phage DNA Isolation Kit (Norgen Biotek Inc., Canada), following the manufacturer's instructions. #46,800) per manufacturer's instructions. You can choose between columns for low throughput . These libraries were pooled and run on a single MiSeq paired-end 300 cycle sequencing run. Imran Sabri. Universidad de Buenos Aires. Genomic DNA. If you are interested in a protocol that's not a comercial kit contact me. A single-end 100-bp high-output sequencing run was conducted using the . Molecular libraries for deep sequencing were prepared using the Nextera XT DNA library preparation kit (Illumina). The final libraries were . As it can be seen, the optional treatment of Proteinase K improved the phage DNA yield in Lanes 2, 5 and 6 . The kit is highly efficient for processing small . Qualitative and quantitative control were performed using conventional electrophoresis and Qubit 2.0 Fluorometer (ThermoFisher Scientific, USA), respectively. The DNA is isolated without the use of phenol, chloroform or cesium chloride. Find phage dna isolation kit and related products for scientific research at MilliporeSigma. The resulting phage particles were then treated with DNase I (ThermoFisher Scientific, Waltham, MA, USA) and RNase I (ThermoFisher Scientific, Waltham, MA, USA) prior to genome DNA extraction using a commercial phage DNA isolation kit (Norgen, Thorold, ON, Canada) and subsequent clean up using a commercial DNA clean up kit (Promega, Madison, WI, USA), used following the manufacturer's . Concentrate the phages by centrifugation at 14,000 rpm for 10 minutes, then proceed with the protocol as written. 46800, 46850) | Norgen Biotek Corp. Phage DNA Isolation Kit (Cat. A kit for the isolation of single strand DNA from M13 phase Cat. Viral Rna Isolation From Blood By Norgen Kit; Stool Dna Isolation Kit; Magmax Rna Isolation Kit; Mircury Rna Isolation Kit (Exiqon; Picopure Rna Isolation Kit Applied Biosystems; Rnaqueous Rna Isolation Kit A1912; Viral Rna Isolation Kit (Sciencell; Viral Rna Isolation Kit; Microbial Dna Isolation Kit; Pureexo Exosome Isolation Kit Norgen Phage Dna Isolation Kits, supplied by Norgen Biotek, used in various techniques. As a result it has excellent search-ability in google and we see a steady increase in interest with a growing list of publications. In particular we have several publications for whole genome . The phage DNA was sequenced on an Illumina MiSeq with 2 150 bp reads, and the genome sequence assembled using Spades v3.11 commercially by SeqMatic (Fremont . DNA extraction using the Norgen Phage DNA Kit This extraction procedure involves several steps but is far simpler than most other methods. Applications Products Services Support. Check out our kits for purifying genomic DNA from a variety of sources, including tissue, blood, plasma, plants, soil, food, formalin-fixed paraffin-embedded (FFPE) samples, forensic samples, and cell lysates. Total DNA was isolated from four enriched phage cultures using Norgen's Phage DNA Isolation Kit. A 'Phage DNA Isolation Kit' (Norgen Biotek, Canada) was used to extract phage DNA isolated from a high titer plate lysate (minimum of 10 8 PFU ml 1. How can I eliminate host bacterial genomic DNA contamination from phage DNA? Cat. P.Iva 10926691006 Via dei Lucani 51, 53, 55 - 00185 Roma Phone: +39.06.9818 5510 Fax: +39.06.8928 0412 Email: info . Norgen's Phage DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. Transcription . Cite. Phage DNA Isolation Kit. The DNA extraction was performed with a phage DNA isolation kit (Norgen Bioteck, Canada). Once extracted, one-step qRT-PCR was performed on 1ul of each sample in duplicate to determine performance between the . The purified nucleic acid was treated with DNase I, RNase A (Thermo Fisher Scientific, USA), and mung bean nuclease (NEB, USA), followed by electrophoresis in a 1% agarose gel to determine the genome type of PVN02. The DNA is isolated without the use of phenol, chloroform or cesium chloride banding procedures. The phage DNA is isolating by norgen DNA isolation kit and the titre is around 10 12. ZERO BIAS - scores, article reviews, protocol conditions and more Here, a lytic E. coli O157:H7-specific phage (KFS-EC) was isolated, identified, and characterized to evaluate its potential as a biocontrol agent for E. coli O157:H7. Five microliter gDNA of each sample was loaded on 1% agarose gel and run for . Phage DNA Isolation Kit by Norgen (Product #26100) 15 mL conical tubes Appropriate pipette tips 96 - 100% Ethanol 96 - 100% Isopropanol Deionized water Proteinase K - 20 mg/mL or other known concentration DNAse I - 2 unit/L or other known concentration HELPFUL TIPS This kit CANNOT BE USED with just ANY lysates. Support by Strong Publication Record . Genomic bacteriophage DNA extraction was performed with a phage DNA isolation kit (Norgen Biotek Corp., Thorold, ON, Canada) according to the manufacturer's instructions. Thermo Fisher Scientific. incubation of the phage . A DNase I pre-treatment (for DNase I kit information see link below) was performed prior to adding the provided Lysis Buffer. Norgen's Phage DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. We recommend preparing buffers, plates, tubes, and media in advance. Bioz Stars score: 99/100, based on 40 PubMed citations. thanks for your answer. Samples were pooled and purified using QIAquick PCR DNA cleanup (Qiagen). This will degrade most of the free DNA leftover from the lysed host cells, but the phage DNA will be protected because it is inside the phage capsid. We started our study by testing five different methods to isolate phage DNA from faecal samples (described in Table 1).These . The whole genome of Vibrio phage . DNA was sequenced by the same protocol as was used for bacterial genome sequencing. The DNA is isolated without the use of phenol, chloroform or cesium chloride. For phage JK08, DNA isolation was performed using the Norgen Phage DNA isolation Kit (Norgen Biotek, ON, Canada) according to the manufacturer's instructions. Methods KFS-EC . Phage DNA was extracted using a phage DNA isolation kit (Norgen Biotek Corp., Ontario, Canada) according to the manufacturer's protocol. Cite . Dear Guanfan Mao, You are requested to please provide the details of procedure you are following, so that we can answer you better. The DNA is isolated without the use of phenol, chloroform or cesium chloride. you can try the Phage DNA Isolation Kit from NORGEN. Nucleic acid was extracted using the MagMAX Viral/Pathogen Nucleic Isolation Kit and and competitor kits from 5 different matrices spiked with inactivated viral particles. Norgen Phage Dna Isolation Kits, supplied by GENEFLOW LIMITED, used in various techniques. Also, the amount of Proteinase K and the incubation time can be increased to improve the DNA yield. Viral Rna Isolation From Blood By Norgen Kit; Stool Dna Isolation Kit; Magmax Rna Isolation Kit; Mircury Rna Isolation Kit (Exiqon; Picopure Rna Isolation Kit Applied Biosystems; Rnaqueous Rna Isolation Kit A1912; Viral Rna Isolation Kit (Sciencell; Viral Rna Isolation Kit; Microbial Dna Isolation Kit; Pureexo Exosome Isolation Kit Dear Guanfan Mao, You are requested to please provide the details of procedure you . To assess the quantity and quality of DNA for sequencing, both a spectrophotometer-based method and Qubit were used. 8th Jan, 2013. Random Amplification of Polymorphic DNA (RAPD) PCR. RAPD PCR was done using DNA Primer P1 (CCGCAGCCAA ) at a concentration of 100 pmol l 1, with 10 l GoTaq Green Mastermix (Promega, UK) and 8 l SDW and added . Products Building Blocks Explorer Technical Documents Site Content Papers Genes. The spin-column based procedure is rapid, and Via dei Lucani 51, 53, 55 Partita Iva 10926691006 Phone: +39.06.9818 5510 - Fax: +39.06.6227 5842 info@aurogene.eu. .. I have two solutions for that: 1) Try to use a pulsed field gel electrophoresis (PFGE), in which your long phage DNA moves out of a well and forms a band; 2) You may use a classical . 27th Sep, 2016. #: DP2301 (50 preps) Signalway Biotechnology . Genome assembly was performed using a BLAST-based de novo assembler, Geneious v10.2.3, and was validated using Unicycler version 0.4.1 . you can try the Phage DNA Isolation Kit from NORGEN. PHAGE GENOMIC DNA EXTRACTION With regard to the safety concerns of using phenol or chloroform and the lack of an ultracentrifuge, we've extracted gDNA of phages by a commercial Phage DNA Isolation Kit 2 (Norgen Biotek Corp.). the procedure we are doing is very simple as describe in the manual of the phage kit. 0 votes 0 thanks. The restriction patterns were visualized . Phage DNA Isolation Kit Norgen Biotek 46850 DNA Purification General; Product specifications for - Phage DNA Isolation Kit . Reducing Phage DNA Yield Phage DNa IsolatIoN KIt NORGEN BIOTEK CORP. From what I've seen most people use either manual extraction, Norgen Biotek's phage DNA isolation kit, or Promega's Wizard DNA clean up system. Phage Dna Isolation Kit, supplied by Norgen Biotek, used in various techniques. Using the Norgen Phage DNA Isolation Kit Finally, centrifugue for at least 2 hours at 16.000 g for . Phage genomic DNA (gDNA) was isolated using Phage DNA Isolation Kit (Norgen Biotek Corp., Ontario, Canada. These fast, easy-to-use kits provide pure, high-quality genomic DNA suitable for such downstream applications as real-time . Pipet fillers ; Bottle-top dispensers ; Micropipettes ; Tips ; bottle-top burettes ; Plate washers . Estefania Urdaniz . #46800: DE3 Lysogenization Kit: Novagen: 69734-3: Materials and equipment. DNA/RNA UV cleaner boxes ; Fume hoods ; View more. Bat Chen R Avraham Lubin. Products; Promotions; Partners; Contacts; Durc; Aurogene s.r.l. Phage DNA Isolation Kit: Norgen Biotek: Cat. Norgen's Phage DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. The genomic DNA was sequenced by Illumina HiSeq sequencing platform and assembled by SPAdes (v3.9.1). Norgen's Phage DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. Whole-genome sequencing was performed at Hokkaido System Science Co., Ltd (Sapporo, Japan) using a high-throughput Illumina HiSeq 2500 system . Sequencing and Bioinformatic Analysis Genome sequencing was conducted by MicrobesNG 1 , which was supported by the BBSRC (grant no. Based on the manufacture's instruction, the protocol was briefly described as follows. Ships Today (13) Product Category . Then, recover the supernatant and you can use chloroform, and RNAse and DNAse for cleaning the sample from hosts acid nucleic. The spin-column based procedure is rapid, and can be completed in less than 45 minutes. A commercially available phage DNA isolation kit (Norgen Biotek Corp, Canada) was used for DNA isolation according to the manufacturer's guidelines. The spin-column based procedure is rapid and can be completed in less than 45 minutes. The numbers within Venn diagrams represent the number of assigned families. King Faisal University. Norgen Biotek: Packing: 50 preps: Price: 291.00 : Expected delivery time: 7 days: Quantity: Spin Up Spin Down. Imran Sabri. Phage genomic DNA was purified using the phage DNA isolation kit (Norgen Biotek) following the recommendations of the manufacturer. Plates could be stored at 4C to avoid contamination. ZERO BIAS - scores, article reviews, protocol conditions and more The whole genome of the phage was sequenced using the Illumina Novaseq 6000 platform with a paired-end library of a read length of 2 150 bp [9] . Figure 2: Viral Extraction Kit Comparison - DNA Virus. Phage DNA Extraction. For DNA analysis 10 L of each 50 L elution was loaded onto a 1X TAE agarose gel and the yield of DNA was compared from the eight different phage types (lane 1 to 8). Spatial Biology; Aurogene s.r.l. Contigs with coverage at least 10 times . Norgen's Phage DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. The spin-column based procedure is rapid and can be completed in less than 45 minutes. Phage DNA Isolation Kit Norgen Biotek 46800 DNA Purification I was wondering if the Qiagen Dneasy blood and . These provide an excellent endorsement of the kit. Ultrapure water systems (Type 1 water) Pure water systems (Type 2 & Type 3 water) Measuring equipment . Proteinase K treatment using Norgen's Phage DNA isolation kit. The Sample Preparation Experts T 1-866-NORGENB T 905-227-8848 F 905-227-1061 E info@norgenbiotek.com www.norgenbiotek.com ISO 13485:2003, ISO 9001:2008, ISO 15189:2012 Registered Product Features Isolate high quality DNA from a broad variety of phage strains High yields of . Isolates DNA From any Bacteriophage, Lambda, M13, T4, T7, Phi a nd any others. Cite. The open reading frames (ORFs) of newly sequenced genomes were predicted with GeneMarkS soft-ware The DNA was resuspended in 50 L of molecular-grade water. Article Title: Characterization and Food Application of the Novel Lytic Phage BECP10: Specifically Recognizes the O-polysaccharide of Escherichia coli O157:H7 The DNA is isolated without the use of phenol, chloroform or cesium chloride. First, DNase is added to the lysates. Bioz Stars score: 86/100, based on 1 PubMed citations. Comments . Electron microscopy was performed at Brigham Young University in the Life Sciences Microscopy Lab using a FEI Helios NATOCAB 600i DualBeam FIB/SEM with STEM detector . BB/L024209/1) for phages UP17, 311F and 05F as well as all the bacterial genomes. 0 votes 0 thanks. Lab water systems . Bioz Stars score: 86/100, based on 1 PubMed citations. The genomic DNA of JK004 was extracted and purified using Phage DNA Isolation Kit (Norgen Biotek, Canada). Consumables/Reagents (See online media preparation guides) Phage DNA Isolation Kit by Norgen (Product #26100) 15 mL conical tubes Appropriate pipette tips 96 - 100% Ethanol 96 - 100% Isopropanol Deionized water Proteinase K - 20 mg/mL or other known concentration DNAse I - 2 unit/L or other known concentration HELPFUL TIPS Purpose Escherichia coli O157:H7 is one of the major foodborne pathogens of global public concern. Advanced Search. The purified . Genomic DNA was extracted and purified from phage particles using a phage isolation kit (Norgen Biotek Corp., ON, Canada) according to the manufacturer's protocol. Bacteriophages (phages) have emerged as a promising alternative to antibiotics for controlling pathogenic bacteria. Treat enriched phage culture with DNaseI. Balances ; Photometers ; Melting point apparatus ; Liquid handling . Norgen Biotek has a Phage DNA Isolation Kit.
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